Unit

UNIT 12.13 Engineering Designer Nucleases with Customized Cleavage Specificities

  1. Jeffry D. Sander1,2,
  2. Morgan L. Maeder1,3,
  3. J. Keith Joung1,2,3

Published Online: 1 OCT 2011

DOI: 10.1002/0471142727.mb1213s96

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Sander, J. D., Maeder, M. L. and Joung, J. K. 2011. Engineering Designer Nucleases with Customized Cleavage Specificities. Current Protocols in Molecular Biology. 96:12.13:12.13.1–12.13.16.

Author Information

  1. 1

    Molecular Pathology Unit, Center for Cancer Research, and Center for Computational and Integrative Biology, Massachusetts General Hospital, Charlestown, Massachusetts

  2. 2

    Department of Pathology, Harvard Medical School, Boston, Massachusetts

  3. 3

    Biological and Biomedical Sciences Program, Harvard Medical School, Boston, Massachusetts

Publication History

  1. Published Online: 1 OCT 2011
  2. Published Print: OCT 2011

Abstract

Engineered designer nucleases can be used to efficiently modify genomic sequence in a wide variety of model organisms and cell types. Zinc finger nucleases (ZFNs), consisting of an engineered zinc finger array fused to a non-specific cleavage domain, have been extensively used to modify a broad range of endogenous genes. Protocols for engineering ZFNs targeted to specific gene sequences of interest using the context-dependent assembly (CoDA) method are described in this unit. Curr. Protoc. Mol. Biol. 96:12.13.1-12.13.16. © 2011 by John Wiley & Sons, Inc.

Keywords:

  • zinc finger nucleases;
  • zinc finger proteins;
  • context-dependent assembly;
  • protein engineering;
  • DNA-binding domains